EFFECT OF HYALURONIC ACID ON CHONDROCYTE RESPONSE TO TUMOR NECROSIS FACTOR-α AND INTERLEUKIN-1

Abstract

Twelve Osteoarthritic human knees(samples obtained at the time of knee joint replacement surgery). I. Animal studies : Chondrocytes were isolated by digestion with pronase and bacterial collagenase, following established procedures, then cultured on alginate beads, grouped as : control, IL-19, TNF-a, HA, IL-19/HA, and TNF-a/HA group. After 48 hours the medium was removed from each well and saved for analysis while the beads were stored at -80°C. II. Human studies: Cartilage slices representing the full thickness of tissue were dissected from human osteoarthritic knee joint, then slices were cultured in a specific mediwn [Ham F-12 + 10% FBSJ for 14-16 hours , then the cartilage slices were divided into 6 groups as: control, IL-13, TNF-, HA, IL-1/HA, and TNF-a/HA. The mediwn was removed from each well after 24 hours {24 hour mediwn samples) and saved at -80C. Then, 3 ml of fresh medium was added to the slices, with the appropriate supplements, and incubation was continued for an additional 96h. The medium was removed (96 hour medium samples) and saved at -80°C. ASSESSMENTS: The following methods were used to assess the efficacy of addition of HA to canine chondrocytes and human cartilage slices in absence and presence of cytokinesIL-1 ~ and TNF-a). 1- Proteoglycan loss into the media, using DMMB binding method in both animal and human studies.